Gene Cloning

Sequencing DNA 205
A7.7. (a) Ten-fold coverage means that on average every base in the genome
has been sequenced 10 times, so you would need to read (4 × 10 6 ) × 10 bases,
i.e. 4 × 10 7 . If the reads are 500 bp you require (4 × 10 7 ) ÷ 500 reads, so the
answer is 80,000.
(b) For the human genome you would need (3 × 10 10 ) ÷ 500 reads, so the
answer is 60 million.
Further Reading
Determination of nucleotide sequences in DNA (1980) Sanger F, From Nobel
Lectures, Chemistry 1971–1980, Editor-in-Charge: Tore Frängsmyr; Editor: Sture
Forsén, World Scientific Publishing Co., Singapore, 1993
This article describes the development of the Sanger dideoxy method of sequencing
DNA. This article can be accessed using the following url:
http://nobelprize.org/chemistry/laureates/1980/sanger-lecture.html
Whole-genome random sequencing and assembly of Haemophilus influenzae Rd
(1995) Fleischmann RD et al., Science, Volume 269 Pages 496–512.
This is a primary research paper describing the sequencing of the first bacterial
genome using a whole-genome shotgun approach
Initial sequencing and analysis of the human genome (2001) International Human
Genome Sequencing Consortium, Nature, Volume 409 Pages 860–921.
The primary research paper reporting the “draft” human genome sequence
Clone by clone by clone (2001) Olson MV, Nature, Volume 409 Pages 816–818.
A commentary on the hierarchical shotgun sequencing approach used to sequence
the human genome
Finishing the euchromatic sequence of the human genome (2004) International
Human Genome Sequencing Consortium, Nature, Volume 431 Pages 931–945.
The primary research paper reporting the “finished” human genome sequence
End of the beginning (2004) Stein LD, Nature, Volume 431 Pages 915–916.
A commentary on the publication of the “finished” human genome sequence
The Sequence of the human genome (2001) Venter JC, et al., Science, Volume 291
Pages 1304–1351.
The primary research paper reporting the “draft” human genome sequence as determined
by the whole-genome shotgun approach
Revisiting the independence of the publicly and privately funded drafts of the
human genome (2003) Cozzarelli NR, Proceedings of the National Academy of
Sciences of the United States of America, Volume 100 Page 3021.
This is an editorial with links to papers that discuss the advantages and disadvantages
of whole-genome shotgun approach to sequencing the human genome
Genome sequencing in microfabricated high-density picolitre reactors (2005)
Margulies M. et al., Nature, Volume 437 Pages 376–380.
The primary research paper discussing 454 pyrosequencing